Superoxide dismutase genes in <i>Pyropia haitanensis</i>: molecular cloning, characterization and mRNA expression

JI Dehua; XU Yan; XIAO Haidong; CHEN Changsheng; XU Kai; XIE Chaotian

doi:10.1007/s13131-016-0873-2

坛紫菜超氧化物歧化酶基因的克隆及表达分析

doi: 10.1007/s13131-016-0873-2

集美大学水产学院, 福建厦门 361021

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- 收稿日期: 2015-03-12
- 修回日期: 2015-08-07

Superoxide dismutase genes in Pyropia haitanensis: molecular cloning, characterization and mRNA expression

Fisheries College, Jimei University, Xiamen 361021, China

摘要

摘要: 超氧化物歧化酶(SOD)是植物细胞应对氧化胁迫的第一道,也是最重要的防线。本研究以坛紫菜转录组数据库中的unigene序列为基础,通过RACE扩增技术克隆得到了3条坛紫菜SOD基因的全长序列,分别命名为PhMSD,PhCSD1和PhCSD2。序列分析结果表明,PhMSD序列全长973 bp,可编码包含224个氨基酸,等电点为5.75的多肽序列;PhCSD1序列全长1029 bp,可编码包含134个氨基酸,等电点为4.65的多肽序列;PhCSD2序列全长954 bp,可编码包含216个氨基酸,等电点为10.74的多肽序列。根据保守序列和系统进化树分析,可以将三条PhSODs分成两个亚型:其中PhMSD属于锰超氧化物歧化酶,PhCSD1和PhCSD2属于铜锌超氧化物歧化酶。采用实时荧光定量PCR技术(qPCR)测定了3条PhSOD基因在坛紫菜不同世代,高温胁迫不同时间水平以及不同失水胁迫梯度下的表达水平变化,结果表明:3条PhSOD基因在叶状体世代中的表达水平均显著高于丝状体世代;在不同水平的高温和失水胁迫下,PhCSD1和PhCSD2的表达水平显著上调,而PhMSD则表现出不同的表达模式,高温胁迫下被显著抑制,失水胁迫下没有发生显著变化。
- 超氧化物歧化酶 /
- 基因克隆 /
- 失水胁迫 /
- 高温胁迫 /
- qPCR
Abstract: Superoxide dismutase (SOD, EC 1.15.1.1) is the first and most important line of cellular defense against oxidative stress. In this study, based on unigene sequences of P. haitanensis, three full-length PhSOD genes were obtained by RACE technology, and named PhMSD, PhCSD1 and PhCSD2. The full-length cDNAs of these genes comprised 973, 1 029 and 954 nucleotides, respectively. The cDNAs encoded proteins of 224, 134 and 216 amino acids, with isoelectric points of 5.75, 4.65 and 10.74, respectively. Based on their conserved motifs and phylogenetic tree analysis, the three PhSODs were divided into two SOD types: PhMSD is a Mn-SOD, and PhCSD1 and PhCSD2 are CuZn-SODs. qPCR was used to measure the expression of the three PhSOD genes in different life phases of P. haitanensis, and under different periods of high-temperature stress and different levels of desiccation. In the different life phases of P. haitanensis, the expression levels of the three PhSODs were all signi.cantly higher in the thallus than in the conchocelis. During high-temperature and desiccation stress, the expression levels of PhCSD1 and PhCSD2 were highly induced by the O₂^- content, but the expression level of PhMSD was highly depressed by high temperature stress and showed no significant change during desiccation.
- superoxide dismutase /
- gene cloning /
- desiccation stress /
- high-temperature stress /
- qPCR

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参考文献(28)

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坛紫菜超氧化物歧化酶基因的克隆及表达分析

doi: 10.1007/s13131-016-0873-2

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Superoxide dismutase genes in Pyropia haitanensis: molecular cloning, characterization and mRNA expression

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